Expression, tissue distribution and subcellular localization of Bm595 from silkworm (Bombyx mori).

Objective In screening the pupa cDNA library, a novel gene Bm595 was identified, the tissue distribution and localization of Bm595 were analyzed in silkworm. Method The recombinant plasmid was transformed into E.coli BL 21 which were induced with IPTG. The purified fusion protein was used to immune a rabbit and the polyclonal antibody was prepared. The tissue distribution of Bm595 was analyzed by real-time PCR and Western blot. Result The quantitative real-time PCR results indicated that the transcription level of Bm595 was the highest in the fifth instar larva and the lowest in egg. The quantity of Bm595 transcription from high to low was testis, silk gland, gut, head, ovary, fat body, epidermis, and air tube, respectively. Western blot results showed that Bm595 was highly expressed in testis, moderately expressed in silk gland and head. Bm595 also existed in fat body. Bm595 was mainly localized in nucleus by immunocytochemistry. Conclusion A preliminary analysis on Bm595 was made at the mRNA, protein expression and cell level which laid a good foundation for the further studies on Bm595 function.